ISH Template Information Form
Incomplete forms may result in a significant delay to the
in situ hybridization.
Shipping Information
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Clearly label each tube with template name, sense/anti-sense and concentration.
If possible, use tubes with screw top caps instead of snap caps in order to prevent leakage during transport.
Contact your sales associate by email or phone prior to shipping materials to Phylogeny.
Include a digital copy of this form in your email and include a hard copy of this form in the shipping package.
Gene templates do NOT need to be cool (wet ice) or frozen (dry ice) during shipment.
Ship probes with a completed copy of this form by FedEx to:
Phylogeny, Inc.
Attention: Jim Williams
1476 Manning Parkway
Powell, OH 43065
Office: (614) 846-6161
Cell: (614) 207-6639
Fax: (877) 591-1815
jim@phylogenyinc.com
Client Information
Investigator Name:
Company / Institution Name:
Mailing Address:
City, State & Zip Code:
Phone:
FAX:
Email Address:
Study ID:
Study Information
Briefly describe study, including numbers and types of tissues and anticipated sites of expression if known:
Template Information
Templates should be prepared in accordance with the “Preparation of Linear Template from Circular Plasmid DNA”
protocol. Templates prepared in other manners will not be accepted unless previously discussed with your sales
associate. Please verify the orientation of templates by sequencing.
PCR Fragments: Ligations of promoters to PCR fragments, such as Ambion's Lig'nScribe Promoter Addition Kit are
also acceptable templates. In this case, simply provide the PCR product size, the polymerases to use for each
template, the DNA concentrations and a picture of the agarose gel with molecular weight markers with sizes
indicated. Send 1 µg of each PCR template in 25 µl TE pH 8.0.
Cloned Genes: Send 10 µg of linearized sense and 10 µg of the linearized antisense templates. Purify the
templates by doing a single phenol/chloroform extraction and a precipitation with 0.3 M sodium acetate and 2
volumes of ethanol. Redissolve to 1 µg/µl in TE pH 8.0 and mail us the templates.
Probe Size: Probes can range from 200 bp to 1.5 kb; probes of 600 to 800 bp are typically very successful.
Template Name:
Vector Containing Template (or PCR):
Probe Template Insert Size (base pairs):
Anti-Sense Template
*Restriction Sites into which cDNA insert was subcloned:
*Restriction enzyme used to generate template:
Polymerase (T3, T7, SP6) to generate the anti-sense cRNA:
Amount (ugs):
Concentration (ug/ul):
Percent GC Content:
Tube Label:
Sense Template
*Restriction Sites into which cDNA insert was subcloned:
*Restriction enzyme used to generate template:
Polymerase (T3, T7, SP6) to generate the sense cRNA:
Amount (ugs):
Concentration (ug/ul):
Percent GC Content:
Tube Label:
Additional Comments (if needed):
* Denotes field that only needs to be completed when submitting circular plasmid DNA.
Linear Gel Image
Please include a picture of a gel of the sense and antisense template restriction digests stained with ethidium
bromide. Please include: molecular weight markers in one lane, undigested plasmid DNA in one lane and
digested DNA in one lane.
Expression Information
Critical to Phylogeny’s in situ hybridization is an understanding of the expression level of
the gene. The most useful information is from a radiometric Northern. If available, please
provide an image of a radiometric northern and indicate the time of exposure. If Northern
data is not available, please provide any information that you have on expression level that
was measured by RT-PCR methods, such as number of cycles or relative expression levels
measured by quantitative PCR. If you have detected the protein by antibody staining or
some other method, please include these details as well.